scholarly journals Diuretics prevent Rho-kinase activation and expression of profibrotic/oxidative genes in the hypertensive aortic wall

2016 ◽  
Vol 10 (6) ◽  
pp. 338-347 ◽  
Author(s):  
Patricio Araos ◽  
David Mondaca ◽  
Jorge E. Jalil ◽  
Cristián Yañez ◽  
Ulises Novoa ◽  
...  
Keyword(s):  
Gene Expression ◽  
Blood Pressure ◽  
Growth Factor ◽  
Aortic Wall ◽  
Transforming Growth Factor ◽  
Rho Kinase ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Tissue Growth ◽  
Hypertensive Rats ◽  
Rock Inhibitor

Background: Diuretics are current antihypertensive drugs since they reduce blood pressure and cardiovascular risk. Increased vascular tone is modulated in a relevant way by the RhoA/Rho-kinase (ROCK) pathway, by acting on vascular smooth muscle cell contraction. This pathway has also proremodeling vascular effects. There are few data on the role of diuretics on both vascular ROCK activation and on proremodeling effects. We assessed the effects of hydrochlorothiazide (HCTZ) and spironolactone (spiro) alone and in combination with the ROCK inhibitor fasudil (FAS) on ROCK activation, gene expression of proremodeling markers and on hypertrophy in the aortic wall of hypertensive rats. Methods: Deoxycorticosterone acetate (DOCA)-salt hypertensive rats (male, Sprague–Dawley) were randomized to the specific ROCK inhibitor FAS, HCTZ, spiro or the combinations of FAS/HCTZ or FAS/spiro for 3 weeks. At the end of the study, ROCK activation (by western blot), gene expression of proremodeling markers (by reverse transcription polymerase chain reaction, RT-PCR) and vascular hypertrophy (by morphometry) were determined in the aortic wall. Results: All treatments significantly reduced blood pressure. In the DOCA rats the p-myosin phosphatase target protein-1 (MYPT1)/t-MYPT1 ratio, index of ROCK activation was higher by 2.8 fold ( p < 0.05) compared with control rats. All treatments reduced ROCK activation in the aortic wall to control levels ( p < 0.05). Besides, significantly increased protein levels of transforming growth factor β1 (TGF-β1), gene expression of TGF-β1, connective tissue growth factor (CTGF), p22 phox and gp91 phox subunits of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, as well as increased media thickness and aortic media area/lumen area (AM/LA) in the untreated hypertensive rats were significantly reduced ( p < 0.05) to control levels by all treatments. Similar effects were observed using both diuretics alone or in combination with FAS. Conclusions: In the aortic wall, both HCTZ and spiro in antihypertensive doses reduce ROCK activation, subsequent expression of genes that promote vascular remodeling and hypertrophy in this experimental model of hypertension. These effects could explain some of their clinical benefits in hypertensive patients.

Uphill running improves rat Achilles tendon tissue mechanical properties and alters gene expression without inducing pathological changes

2012 ◽  
Vol 113 (5) ◽  
pp. 827-836 ◽  
Author(s):  
K. M. Heinemeier ◽  
D. Skovgaard ◽  
M. L. Bayer ◽  
K. Qvortrup ◽  
A. Kjaer ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mechanical Properties ◽  
Growth Factor ◽  
Achilles Tendon ◽  
Glucose Uptake ◽  
Transforming Growth Factor ◽  
Achilles Tendinopathy ◽  
Tissue Growth ◽  
Tendon Tissue ◽  
Collagen Iii

Overuse Achilles tendinopathy is a common and challenging problem in sports medicine. Little is known about the etiology of this disorder, and the development of a good animal model for overuse tendinopathy is essential for advancing insight into the disease mechanisms. Our aim was to test a previously proposed rat model for Achilles tendon overuse. Ten adult male Sprague-Dawley rats ran on a treadmill with 10° incline, 1 h/day, 5 days/wk (17–20 m/min) for 12 wk and were compared with 12 control rats. Histological, mechanical, and gene-expression changes were measured on the Achilles tendons after the intervention, and local tendon glucose-uptake was measured before and after the intervention with positron emission tomography. No differences were detected between runners and controls in tissue histology or in glucose uptake, indicating that tendon pathology was not induced. Greater tendon tissue modulus ( P < 0.005) and failure stress/body weight ( P < 0.02) in runners compared with controls further supported that tendons successfully adapted to uphill running. Several genes of interest were regulated after 12 wk of running. Expression of collagen III and insulin-like growth factor I was increased, while collagen I was unchanged, and decreases were seen in noncollagen matrix components (fibromodulin and biglycan), matrix degrading enzymes, transforming growth factor-β1, and connective tissue growth factor. In conclusion, the tested model could not be validated as a model for Achilles tendinopathy, as the rats were able to adapt to 12 wk of uphill running without any signs of tendinopathy. Improved mechanical properties were observed, as well as changes in gene-expression that were distinctly different from what is seen in tendinopathy and in response to short-term tendon loading.

Characterization of connective tissue growth factor expression in primary cultures of human tubular epithelial cells: modulation by hypoxia

2010 ◽  
Vol 298 (3) ◽  
pp. F796-F806 ◽  
Author(s):  
Sven Kroening ◽  
Emily Neubauer ◽  
Bernd Wullich ◽  
Jan Aten ◽  
Margarete Goppelt-Struebe
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Epithelial Cells ◽  
Connective Tissue ◽  
Connective Tissue Growth Factor ◽  
Transforming Growth Factor ◽  
Primary Cultures ◽  
Tissue Growth ◽  
Tubular Epithelial Cells ◽  
Mesenchymal Phenotype

Tubular epithelial cells secrete connective tissue growth factor (CTGF, CCN2), which contributes to tubulointerstitial fibrosis. However, the molecular regulation of CTGF in human primary tubular epithelial cells (hPTECs) is not well defined. Therefore, CTGF expression was characterized in hPTECs isolated from healthy parts of tumor nephrectomies, with special emphasis on the regulation by transforming growth factor-β (TGF-β) and hypoxia, essential factors in the development of fibrosis. CTGF synthesis was strongly dependent on cell density. High CTGF levels were detected in sparse cells, whereas CTGF expression was reduced in confluent cells. Concomitantly, stimulation of CTGF by TGF-β or the histone deacetylase inhibitor trichostatin was prevented in dense cells. Exposure of hPTECs to low oxygen tension (1% O2) or the hypoxia mimetic dimethyl-oxalylglycine for 24 h reduced CTGF gene expression in most of the 17 preparations analyzed. Preincubation of the cells under hypoxic conditions significantly reduced TGF-β-mediated upregulation of CTGF. In line with these data, CTGF mRNA was only induced in interstitial cells, but not in tubular cells in kidneys of mice exposed to hypoxia. Longer exposure to hypoxia or TGF-β (up to 72 h) did not induce hPTECs to adopt a mesenchymal phenotype characterized by upregulation of α-smooth muscle actin, downregulation of E-cadherin, or increased sensitivity of the cells in terms of CTGF expression. Sensitivity was restored by inhibition of DNA methylation. Taken together, our data provide evidence that exposure to hypoxia decreased CTGF gene expression. Furthermore, hypoxia per se was not sufficient to induce a mesenchymal phenotype in primary tubular epithelial cells.

scholarly journals Global Transcriptomic Analyses Reveal Genes Involved in Conceptus Development During the Implantation Stages in Pigs

2021 ◽  
Vol 12 ◽  
Author(s):  
Xupeng Zang ◽  
Ting Gu ◽  
Qun Hu ◽  
Zhiqian Xu ◽  
Yanshe Xie ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Heart Development ◽  
Molecular Mechanisms ◽  
Transforming Growth Factor ◽  
Fetal Loss ◽  
Tissue Growth ◽  
Morphological Development ◽  
Aaa Atpase ◽  
Conceptus Development

Prenatal mortality remains a significant concern to the pig farming industry around the world. Spontaneous fetal loss ranging from 20 to 45% by term occur after fertilization, with most of the loss happening during the implantation period. Since the factors regulating the high mortality rates of early conceptus during implantation phases are poorly understood, we sought to analyze the overall gene expression changes during this period, and identify the molecular mechanisms involved in conceptus development. This work employed Illumina’s next-generation sequencing (RNA-Seq) and quantitative real-time PCR to analyze differentially expressed genes (DEGs). Soft clustering was subsequently used for the cluster analysis of gene expression. We identified 8236 DEGs in porcine conceptus at day 9, 12, and 15 of pregnancy. Annotation analysis of these genes revealed rRNA processing (GO:0006364), cell adhesion (GO:1904874), and heart development (GO:0007507), as the most significantly enriched biological processes at day 9, 12, and 15 of pregnancy, respectively. In addition, we found various genes, such as T-complex 1, RuvB-like AAA ATPase 2, connective tissue growth factor, integrins, interferon gamma, SLA-1, chemokine ligand 9, PAG-2, transforming growth factor beta receptor 1, and Annexin A2, that play essential roles in conceptus morphological development and implantation in pigs. Furthermore, we investigated the function of PAG-2 in vitro and found that PAG-2 can inhibit trophoblast cell proliferation and migration. Our analysis provides a valuable resource for understanding the mechanisms of conceptus development and implantation in pigs.

scholarly journals Ocular surface cytokine profile in chronic Stevens-Johnson syndrome and its response to mucous membrane grafting for lid margin keratinisation

2017 ◽  
Vol 102 (2) ◽  
pp. 169-176 ◽  
Author(s):  
Srividya Gurumurthy ◽  
Geetha Iyer ◽  
Bhaskar Srinivasan ◽  
Shweta Agarwal ◽  
Narayanasamy Angayarkanni
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Mucous Membrane ◽  
Ocular Surface ◽  
Transforming Growth Factor ◽  
Tissue Growth ◽  
Stevens Johnson Syndrome ◽  
Gm Csf ◽  
Johnson Syndrome ◽  
Collagen Iii

BackgroundTo study the tear cytokine and the conjunctival and oral mucosal marker profile in chronic ocular Stevens-Johnson syndrome (SJS) and their alteration following mucous membrane grafting (MMG) for lid margin keratinisation (LMK).MethodsIn a 1-year prospective study, SJS cases (n=25) and age-matched/sex-matched healthy controls (n=25) were recruited. Tear specimen (Schirmer’s strip), conjunctival and oral mucosal imprints were collected from controls and SJS cases pre-MMG and post-MMG (at first follow-up, n=17). Tear cytokines were profiled using 27-bioplex array. Transforming growth factor-beta (TGF-β)-mediated extracellular matrix changes in conjunctival and oral mucosal cells were analysed by gene expression studies. 30ResultsTear cytokine profiling of chronic SJS cases at pre-MMG stage revealed significant upregulation of cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-8, IL-1β, monocyte chemoattractant protein-1, IL-15, IL-2, IL-17A and basic fibroblast growth factor (bFGF) with downregulation of IP-10 (interferon gamma-induced protein 10), tumour necrosis factor-α, interferon-γ, IL-10, vascular endothelial growth factor, regulated upon activation normal T-cell expressed and secreted (RANTES), IL-7, IL-12p70 and IL-13, with maximal increase in GM-CSF and maximal downregulation of IP-10, respectively. Of these, IL-2, IL-15, bFGF and IL-17A showed significant correlation with disease severity, pre-MMG. Conjunctival cells pre-MMG showed increase in TGF-β1, TGF-βRII, connective tissue growth factor and collagen-III gene expression by 10, 67, 173 and 184 folds, respectively, which dropped to 1.3, 11, 13.5 and 19 folds correspondingly, post-MMG. However, their expressions in oral mucosa were negligible.ConclusionA proinflammatory, profibrotic, antiapoptotic ocular surface milieu characterises chronic ocular SJS. IP-10, an antifibrotic cytokine was noted to be maximally downregulated, unlike in other forms of chronic dry eye disease. The alterations in the ocular surface are seen to reverse largely with MMG for LMK.

scholarly journals Transforming Growth Factor-β Gene Expression Studies in Nasal Mucosal Biopsies in Naturally Occurring Allergic Rhinitis

2007 ◽  
Vol 89 (6) ◽  
pp. 563-573 ◽  
Author(s):  
Rami J Salib
Keyword(s):  
Gene Expression ◽  
Allergic Rhinitis ◽  
Growth Factor ◽  
Mast Cells ◽  
Inflammatory Response ◽  
Seasonal Allergic Rhinitis ◽  
Transforming Growth Factor ◽  
Tissue Growth ◽  
Naturally Occurring ◽  
Tnf Α

Introduction Evidence has been provided of enhanced epithelial transforming growth factor-beta (TGF-β) immunoreactivity in allergic rhinitis, including correlation with intra-epithelial mast cell numbers, and the co-localisation of TGF-β receptors to mast cells, suggesting that the epithelial expression of TGF-β may represent an important biological process involved in either the recruitment or retention of mast cells within the epithelium in naturally occurring allergic rhinitis. Patients and Methods In order to extend the above findings, evaluation was undertaken in whole nasal biopsies from subjects with naturally occurring allergic rhinitis, of levels of TGF-β isotypes and receptors gene expression using real-time quantitative polymerase chain reaction (TaqMan RT-PCR), and the results compared to those for tumour necrosis factor-alpha (TNF-α), as a positive control. The study was also extended to evaluate gene expression for connective tissue growth factor (CTGF) and Smad proteins, as downstream markers of TGF-β bioactivity, in the same populations. Results There were no significant differences between the rhinitic and non-rhinitic groups in the expression of TGF-β isoforms or Smad-3, Smad-6 and Smad-7 proteins; however, there was increased gene expression for TGF-βRI and TGF-βRII along with CTGF in seasonal allergic rhinitis. TNF-α gene expression was also increased in seasonal allergic rhinitis, consistent with a more acute inflammatory response in this form of rhinitis. Conclusions This study advances our understanding of the role of TGF-β in the pathogenesis of the inflammatory response in allergic rhinitis.

scholarly journals Ethyl Acetate Fraction of Amomum xanthioides Exerts Antihepatofibrotic Actions via the Regulation of Fibrogenic Cytokines in a Dimethylnitrosamine-Induced Rat Model

2016 ◽  
Vol 2016 ◽  
pp. 1-14 ◽  
Author(s):  
Sung-Bae Lee ◽  
Hyeong-Geug Kim ◽  
Hyo-Seon Kim ◽  
Jin-Seok Lee ◽  
Hwi-Jin Im ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Liver Fibrosis ◽  
Ethyl Acetate ◽  
Rat Model ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Ethyl Acetate Fraction ◽  
Tissue Growth ◽  
Growth Factor Beta

Amomum xanthioides has been traditionally used to treat diverse digestive system disorders in the Asian countries. We investigated antihepatofibrotic effects of ethyl acetate fraction of Amomum xanthioides (EFAX). Liver fibrosis is induced by dimethylnitrosamine (DMN) injection (intraperitoneally, 10 mg/kg of DMN for 4 weeks to Sprague-Dawley rats). EFAX (25 or 50 mg/kg), silymarin (50 mg/kg), or distilled water was orally administered every day. The DMN injection drastically altered body and organ mass, serum biochemistry, and platelet count, while EFAX treatment significantly attenuated this alteration. Severe liver fibrosis is determined by trichrome staining and measurement of hydroxyproline contents. EFAX treatment significantly attenuated these symptoms as well as the increase in oxidative by-products of lipid and protein metabolism in liver tissues. DMN induced a dramatic activation of hepatic stellate cells and increases in the levels of protein and gene expression of transforming growth factor-beta (TGF-β), platelet derived growth factor-beta (PDGF-β), and connective tissue growth factor (CTGF). Immunohistochemical analyses revealed increases in the levels of protein and gene expression of α-smooth muscle actin. These alterations were significantly normalized by EFAX treatment. Our findings demonstrate the potent antihepatofibrotic properties of EFAX via modulation of fibrogenic cytokines, especially TGF-β in the liver fibrosis rat model.

Sign in / Sign up

Export Citation Format

Share Document